An easy and precise tool for the early detection of malignancies is required for rapid care and metastasis prevention. Cobalt-based metal–organic framework (Co-BTC-GO-MOF) nanoparticles have been used as a signal carrier for the anti-VEGF165 signaling antibody. Cobalt-based MOF was synthesized using cobalt (Co), benzene-1,3,5-tricarboxylate (BTC), and graphene oxide (GO) applying a hydrothermal method. Structure, compositions, size and morphology of the qualified sensor are determined by using distinctive analytical techniques. The Co-MOF nanoparticles are found to be thermostable, as revealed by thermal stability assay. The strategy utilises an impedimetric and differential pulse voltammetry (DPV) techniques in the presence of the [Fe(CN)6]3−/4− redox system. Compared to earlier results, this assay resulted in higher sensitivity with the limit of detection (LOD) found to be 5.23 pM in a 0.01 M buffer solution of pH 7.4 using linear scale voltammetry at room temperature. The resulting Co-BTC-GO-MOF immunosensor shows high responsiveness and selectivity in detecting VEGF165 in real-time serum samples of cancer patients. The electrochemical performance studies confirm that the intended proposed immunosensor could pave the way for the future advancement of high-performance, sensitive, reproducible and robust immunosensors for the cost-effective and initial phase detection of cancer in the future.The vascular endothelial growth factor 165 (VEGF 165) is a quintessential biomarker in cancers.The vascular endothelial growth factor 165 (VEGF An abnormal level (either over expression or down-regulation in the level of VEGF) is responsible for the induction of various diseases. The over expression represents a potent promoter in the pathophysiology of angiogenesis associated with the growth of tumors. VEGF is overexpressed in numerous human cancers like lung, breast, brain cancer, urinary tract and gastrointestinal cancers.2 Alternatively, when the level of VEGF is lower, it is an indication of developing degeneration of neurons by limiting the neural tissue perfusion. Different CNS disorders, such as Parkinson's disease and brain injuries, are associated with the down-regulation of VEGF.3 Some major expressed variants of VEGF isoforms from a single VEGF gene consist of 121, 145, 165, 183, 189 and 206 amino acids. Out of these six isoforms, VEGF165 is the most predominant VEGF-A isoform, which is over expressed in tumor cells during preliminary tumor growth phase, especially, in breast and lung cancers and possible predictor of cancers. Therefore, VEGF165 is an important predictive biomarker for early detection of cancer in healthcare settings. The determination of the level of VEGF165 in blood, irrespective of their type, can be a fruitful approach to clinical diagnosis of cancer.4Vascular endothelial growth factor (VEGF) is a hypoxia-inducible protein that results from alternate splicing of the VEGF gene.165, which include enzyme-linked immunosorbent assay (ELISA),5 optical methods,6 electrochemical immunoassay methods7 and radioimmunoassay techniques.8 The ELISA continues to be the gold standard for clinical quantification of many protein biomarkers with excellent specificity and very low limits of detection (LOD).9 However, the requirement of sophisticated instrumentations, complex detection protocols with a long testing time, lack of portability, difficulty with multiplexed sensing and high cost, it is not suitable for point-of-care applications.10 To overcome these shortcomings of the existing detection techniques, the emerging electrochemical immunosensor has inherent benefits. As compared to traditional molecular recognition system selector chemical immunosensor is widely accepted because of its simple instrumentation, high sensitivity, affinity, portability, rapidity, reproducibility and wide applicability at an economical price. They are also regarded as important tools for monitoring and preventing various diseases, including cancers. On the other hand, due to the high specificity of the antibody towards specific antigen and the faster reaction time, the antibody can be explored as a potential tool in detecting substances.11 In consequence, it is highly desirable to design a novel nanomaterial for selective and sensitive detection of VEGF165 even at lower concentrations.Till date, there are several methods have been reported for detection and quantification of VEGF12 Consequently, MOFs can serve as an excellent platform to effectively ligate the recognized element to the surface of the electrode, which leads to adequate signal amplification.In the past decade, metal–organic frameworks (MOFs) based nanomaterials have emerged as an excellent alternative tool for quantification of biomarkers. It has been reported to be a very useful tool to modify electrodes due to its highly selective reactivity, high yield, moderate reaction conditions and simplicity. On top of that, the electrochemical application of MOFs in recent years has been fascinated due to its large pore size, high specific surface area, and good conductivity. Additionally, the specific antibodies can be incorporated in the architecture of MOFs by covalent bonding.13 However, the low dispersibility, poor electronic conductivity and limited sensitivity of MOFs is still a challenge.14 The addition of graphene oxide (GO) can mitigate these problems due to its unique structure and distinguished physiochemical and mechanical properties. It can produce a synergetic effect by increasing the dispersibility, subsequently electronic conductivity and sensitivity of the composite.15 Based on these advantages, in the present work, for the first time, cobalt (Co), benzene-1,3,5-tricarboxylate (BTC) and GO-based MOFs (Co-BTC-GO-MOFs) were synthesized by a simple hydrothermal method for detection of VEGF165 on both, signal on impedimetric and differential pulse voltammetry (DPV) techniques. The use of CO-BTC-MOFs combined with GO yielded improved sensitivity and stability. Moreover, the signal readout was also much simpler and low-cost with the target to provide good sensing performance, such as fast reaction, high sensitivity, with good stability and reproducibility.In the present work, nano-porous MOF is designed. There are several electrochemically active MOFs have been introduced as an electrocatalyst, electroactive signal probes or redox-active species, such as Co-MOF, Ni-MOF and Cu-MOF, for detection of various analytes.2165, anti-VEGF165 antibody, N-hydroxysuccinimide (NHS), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), dimethylformamide (DMF) and ethanol (95%) were purchased from Sigma-Aldrich, India. Just to highlight that anti-VEGF antibody and VEGF165 were stored at 4 °C and −20 °C, respectively, before use. Freshly aqueous solutions of EDC and NHS were prepared before using. All solutions were prepared using deionized water (DI) from a Millipore system. Cobaltous chloride, potassium ferricyanide, 1,3,5-benzenetricarboxylic acid (BTC) and GO were purchased from Merck. Sodium dihydrogen orthophosphate dehydrate (Merck) and sodium phosphate dibasic dehydrate (Sigma-Aldrich) were used to prepare a 0.1 M phosphate buffer of different pH to carry out pH study. All chemicals were of analytical grade and used without any further purification.Human recombinant VEGF2.1α X-ray radiation (λ = 1.5418 Å) at a scan rate of 0.02 s−1, which is operated at 40 kV and 50 mA. Fourier transform infrared spectroscopy  and thermal stability of the prepared samples were recorded using Q Series Thermal Analyzer DSC/TGA (Q600). The Brunauer–Emmett–Teller (BET) specific surface area and porosity were recorded using Micromeritics Tristar II ASAP 2020 system, USA surface area and porosity analyser.Electrochemical measurements were performed on a three-electrode cell system of the electrochemical workstation  using glassy carbon electrode (GCE – 3 mm diameter) as working, Ag/AgCl, as reference and platinum wire as a counter electrode, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) was performed in a potential window of −0.4 to 0.6 V in 0.1 M PBS at room temperature. Morphology and surface of the Co-BTC-GO-MOFs was observed under the field emission-scanning electron microscope . The Co-BTC-GO-MOFs were sputter-coated with gold to avoid accumulation of charges before FE-SEM examination. For texture analysis and imaging the sample holder was then placed into the FE-SEM. Observation magnification may be regulated and considered based on the requirements. Further, high resolution-transmission electron microscopy  were used for the determination of size. It is also helpful in providing direct images of the atomic structure of the Co-BTC-GO-MOFs. The Co-BTC-GO-MOFs were dissolved in DI water then mounted onto the surface of a copper grid. The excessive sample was removed by blotting off with filter paper, then dried at room temperature before analysis. X-ray diffraction patterns (XRD), were recorded using Philips X'Pert, UK fitted with Cu-K2.216Co-BTC-GO-MOFs were synthesized by mixing 0.535 mg of cobaltous chloride (2.25 mmol) dissolved in 7.5 mL of DI water and 0.264 g of BTC (0.264 mmol) dissolved in 7.5 mL of ethanol. The mixture was kept under constant magnetic stirring for 10 min. A 2 mg of GO was added in the above mixture under constant stirring, and the whole mixture is in Teflon lined stainless steel autoclave (200 mL), which is subjected to hydrothermal reaction at 100 °C for 24 hours. The precipitate was then washed with a copious amount of DI water to remove the excess reagents using centrifuged at 4000 rpm. The finished final product was obtained by centrifugation, washed several times with DI water and kept for vacuum drying to achieve constant weight at room temperature. Subsequently, for further use, the lyophilised sample was kept in airtight glass vials.2SO4 solution and polished with α-alumina powder  on a polishing cloth to achieve a mirror like surface. Further, it was again ultrasonically cleaned in the solution mixture of DI water and ethanol. It was finally dried with acetone.Before proceeding to fabricate the developed Co-BTC-GO-MOFs, the surface of the bare GCE was electrochemically clean in 0.1 M H−1 of dispersion of Co-BTC-GO-MOFs was prepared in DI water. In order to prepare an electrode, 5 μL dispersion mixture of Co-BTC-GO-MOFs was dropped onto the dry and clean GCE surface. It was kept for drying under an infrared lamp. Following the drying process, the electrode was then activated by the treatment of EDC/NHS (30 mM/2 mM) for 2 h. It was subsequently washed with DI water. Further, the electrode was incubated in VEGF165 antibody solution (10 μg mL−1) prepared in 0.1 M PBS, pH 7.4. It was immersed for 4 h. Finally, the modified electrode was washed with 0.1 M PBS and concurrently blocked with bovine serum albumin (BSA) solution  to eliminate the non-specific binding effects. It was followed by careful washing in 10 mM PBS (pH 7.4). Thus, the fabricated immunosensor was obtained.Subsequently, 1 mg mL165 antigen were freshly prepared in PBS by serial dilution from the stock solution. The prepared antigen solutions were kept on ice throughout the experiments. 5 μL of each antibody solution was drop casted on the biosensor and incubated at room temperature for 1 hour. After that, the sensor was rinsed with PBS to remove any unbound peptides. The measurement time for each sensor is approximately 3–4 minutes. Here, the DPV technique was adopted to measure the responsive signals from redox probes. The variable experiment parameters were set for the current measurements are initial potential: −0.8 V; final potential: 0.4 V; pulse amplitude: 0.05 V; pulse width: 0.25 s; sample interval: 0.002 s.Once the antibody-modified sensor was obtained, different concentrations of solution (5 μL) were introduced onto the surface of the developed electrode. The desired dilution of VEGF165 antigen were prepared in serum by mixing with PBS by using serial dilution technique. Then, biosensors were incubated with spiked samples for 60 min followed by washing with PBS and electrochemical measurement were recorded.Serum sample analysis was carried out for the newly developed immunosensor to check the its practical application and feasibility. We perform the real-sample analysis in sheep blood sample. Sheep blood was obtained from Rockland Immunochemicals, Inc. Blood sample of sheep was diluted with PBS solution in the ratio 1 : 100. The desired dilutions of the VEGF3−1 in both Co-BTC  scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg>O present in the structures, while at about 1245 cm−1 was assigned to C–O. At the fingerprint region, the peak between 600–690 was linked to the Co–O bond.To explore the functional groups, present in the prepared MOFs, FTIR analysis was carried out. All the MOFs composites , and antibody-Co-BTC-GO (ab-Co-BTC-GO) showed similar spectra with exceptions of emerging peaks for composites with graphene oxide and antibody or both. The sharp peak at about 3398 cmh Co-BTC  and Co-Bh Co-BTC  is attriλ = 2d sin θ. The diffraction peak at 2θ = 15° (indexed 001) is attributed to the GO with an interlayer spacing of 1.38 Å, indicating the presence of GO in prepared MOFs. The other main characteristic peaks were observed at 2θ = 19, 30, 31, 37, 40 and 63°, which correspond to (440), (220), (100), (011), (400) and (511) crystal planes cobaltous chloride, 1,3,5-BTC, respectively. This results was in a good agreement with what has been reported in previous works.17The crystal structure of the Co-BTC-GO-MOFs was explored using XRD, as shown in 18–20 To visualize the inner microstructure of the Co-BTC-GO-MOFs, TEM was employed. TEM images . Thus, the second loss of weight for the composite Co-BTC-GO-MOFs was recorded at 384 °C, which resulted in a 60% loss of weight. This could further be explained by the improved thermo-stability of the developed Co-BTC-GO-MOFs as shown in The thermal stability of the formulated composite was studied using TGA analysis at the temperature range between 25–800 °C. Initial loss of weight of the Co-BTC-GO-MOFs during initial sections of TGA curves could be correlated with the evaporation of coordinated water from the complex molecule below 100 °C as shown in 6]3−/4− in 1 M KCl as the electrolyte. CVs of [Fe(CN)6]3−/4− redox couple is a suitable probe to analyze the surface properties of the electrode during numerous modification steps.22 Later, the electrochemical surface area of the modified electrode was calculated using the Randles–Sevcik Ip is the peak current in ampere, A represents the surface area of the electrode (cm2), D is the diffusion coefficient which is 7.60 × 10−6 cm2 s−1 for K3[Fe(CN)6], n represents the number of electrons transferred, which in case of K3[Fe(CN)6] is equal to 1, ν is the scan rate (V s−1) and C0 is the concentration in mol cm−3. The slope of plot Ipavs. ν1/2 in 2 and 0.05877 cm2, 0.09667 cm2, respectively. The large active surface area of Co-BTC-GO/GCE also helps in the immobilization of a higher quantity of antibody on the modified GCE, leading to better detection of VEGF165.The CVs were performed to carry out the electrochemical characterization of the developed Co-BTC-GO-MOFs modified GCE. This CV curve was recorded executed in 1 mM 3−/4− in 0.01 M PBS as the redox couple. The pH value was kept 7.4 as the biological protein get destroy in highly acidic or alkaline conditions. Under a neutral environment, smaller electrostatic repelling is favourable to immune reaction. Each step of fabrication of the sensor electrode was monitored by the electrochemical performance through measurement of DPV. DPV of Co-BTC-GO-MOFs displayed a peak current at the potential of 0.45 V as shown in The experimental conditions were optimized to obtain the optimal electrochemical response during analysis. It is very well-known that the concentration of antibody required for immobilization and BSA incubation time greatly affects the sensitivity, specificity and sensor performance. Therefore, we investigated the concentration of antibody required for immobilization and the BSA incubation time to block all non-specific binding sites effectively. In this study, DPV was performed to analyse the analytical performance and detection capability of the electrochemical immunosensor. The DPV measurements were carried out in 1 mM [Fe(CN)Rct) value of the modified electrode did not increase on increasing the antibody concentration above 0.6 mM observed . This caThe LOD for the fabricated electrode was calculated from the calibration plot, which is found to be 5.23 pM (using relation 3S/N). The present work provided better LOD than the previously reported sensor. The performance comparison of the current sensor with reported sensors in the literature is summarized in 165 in three different serum sample dilutions. The precision of the Co-BTC-GO-MOFs was accessed by adding standard solutions of different concentrations. −13 M to 10−6 M. The linear regression equation came out from The VEGF plays an important role in the development of various tumors and metastasis. Therefore, to investigate the feasibility and potential analytical applicability of the fabricated sensor, the developed immunosensor was assessed through the recognition of VEGF165 in the practical serum specimens from the cancer patients as an approach with sensitivity and viability. Therefore, the proposed Co-BTC-GO-MOFs have a great potential for clinic diagnosis of cancer at the early stage for proper treatment and cure of the patient.From DPV results; it can be predicted that the Co-BTC-GO-MOFs could be a promising tool for determination of VEGF31 Thus, three modified electrodes were independently prepared to evaluate the reproducibility of the proposed Co-BTC-GO-MOFs to assess at the optimized experimental conditions. The obtained results were used to determine the mean relative standard deviation (RSD), where the currents of the obtained DPV signals of the proposed Co-BTC-GO-MOFs based immunosensor was found to be 2.54%. These results clearly indicated that the developed electrodes are highly reproducible. Additionally, the low % RSD values with the Co-BTC-GO-MOFs represents the efficiency of the fabricated electrode for the detection of VEGF165.The reproducibility of scientific assays is an important parameter that needs to be considered in order to develop a reliable sensor.To investigate the stability of the Co-BTC-GO-MOFs based immunosensor, the modified electrode was kept in PBS with a pH of 7.4 at room temperature under ambient for 1, 3, 6, 10, 14 and 30 days. The response of the stored electrode was 99.8, 99.6, 99.5, 99.03, 98.09 and 98.02%, respectively. The response of the modified electrode showed excellent storage stability.4165 using Co-BTC-GO-MOFs as the biosensing platform. The developed immunosensor exhibits a wider linear range with lower detection limit, better anti-interference as well as higher stability and repeatability to the VEGF165 under the optimized experimental parameters. The LOD was found to be 5.23 pM with a linear range of 10−13 to 10−6 M. This result showed that Co-BTC-GO-MOFs could be applied in point of care scenarios. Collectively, these findings shed light on the modern electrochemical analysis platform, which gives us great potential for portable on-site detection of VEGF165, based on this proof-of-concept sensor. Moreover, how these advances can become clinically useful tools. In view of an excellent LOD with a wide linear range, this newly synthesized Co-BTC-GO-MOFs represents an insightful paradigm for effective cancer detection, which could be translated to the clinics.In this study, a novel and ultrasensitive immunosensor has been successfully developed for the detection of VEGFSima Singh – conceptualization, synthesis, characterization, detection and drafting of manuscript; Arshid Numan – conceptualization, electrochemical detection, reviewing and editing; Yiqiang Zhan – fund for the project, supervision, reviewing and editing; Vijender Singh – reviewing of manuscript; Aftab Alam – drafting and reviewing of manuscript; Tran Van Hung – data curation, original draft and editing; Nguyen Dang Nam – supervision, reviewing and final proof read.There is no conflict of interest and disclosures associated with the manuscript.


The MIPs@EDA-CDs/CF could be stored at 4 °C for a few weeks and could be reused twice. The proposed method could apply for the specific determination of MT in water and sediment samples along with satisfactory recoveries of 96–104% and an acceptable relative standard deviation of 1–6% at the ppb level.Molecularly imprinted polymers@ethylenediamine-modified carbon dots grafted on cotton fabrics (MIPs@EDA-CDs/CF) and smartphone-based fluorescence image analysis were proposed and used for the first time for the detection of 17 α-methyltestosterone (MT). The EDA-CDs were synthesized and grafted on cotton fabric before coating with the MIPs. The MIPs were synthesized using the MT as a template molecule, methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a cross-linker, and azobisisobutyronitrile (AIBN) as an initiator. The MIPs@EDA-CDs/CF were characterized using FTIR, SEM-EDS, and RGB fluorescence imaging. The fluorescence images were also taken using a smartphone and the ImageJ program was used for RGB measurement. The Δ red intensity was linearly proportional to MT concentration in the range of 100 to 1000 μg/L (R Endocrine-disrupting chemicals (EDCs) have been released into the environment in large numbers and quantities since World War II. EDCs are substances or chemical mixtures that interfere with the function of the human body’s hormones . EDCs arMT, a synthetic hormone that binds to the androgen receptor, is known for interfering effects on the endocrine systems of invertebrate and vertebrate organisms . It is aSeveral methods combined with analytical instruments including gas chromatography–mass spectrometry (GC–MS) , high-peCotton fabrics, a renewable bioresource, are primarily made up of the most abundant biopolymer cellulose with some noncellulosic components surrounding the cellulose core, such as hydrophobic impurities  ,25. TheyCDs have received much attention in recent decades. These particles are synthesized from carbon materials and are one of the newest members of the quantum nanomaterials family. Since the discovery of CDs, these nanomaterials have received a lot of attention because of their superior properties, including easy functionalization, good photostability, excellent water solubility, bright fluorescence emission, tunable surface functionalities, great and green biocompatibility, low toxicity, and easy synthetic routes ,29 with Molecular imprinting technology is based on the principle of enzymes and substrates, hormones and receptors, and antibodies and antigens, which are specific to the recognition of the target analytes . MIPs caSeveral techniques of synthesizing MIPs for target analytes have been reported ,45,46,47ex) was found at 353 nm, and the maximum fluorescence emission wavelength (λem) was found at 450 nm. The exposure time for synthesizing EDA-CDs through the microwave was investigated at a fix power of 300 W. The fluorescence emission spectra of 1000 mg/L EDA-CDs solution from different exposure times (λex = 353 nm) is shown in A 1000 mg/L EDA-CDs solution  was analyzed using a fluorescence spectrophotometer to obtain the excitation spectrum and fluorescence emission spectrum, as shown in As shown in −1 (32 scans and 2 resolutions). Attribution to the vibrational modes of citric acid molecules in −1, and 1397 cm−1 confirmed the presence of the -OH group. In addition, the peak at 1731 cm−1 was observed, which was attributed to the C=O stretching of the carboxylic group of CA while C-OH stretching and CH2 rocking vibrations were observed at 1078 cm−1 and 790 cm−1, respectively. Compared with unmodified CDs, the peak C=O stretching at 1731 cm−1 disappeared from the EDA-CDs spectrum as shown in −1 and N-H bending vibrations at 1558 cm−1, in addition to C-NH-C asymmetric stretching at 1118 cm−1, confirmed the presence of the CONH groups on the CDs [As shown in  the CDs .The adsorption ability of the EDA-CDs/CF was investigated by measuring the differences between the initial concentrations and remaining concentrations of EDA-CDs in a solution using a calibration method from a fluorescence spectrophotometer. To obtain the highest adsorption, the volume of EDA-CDs solutions (1000 mg/L) and sonication times were optimized using the volume from 5 to 20 mL and sonication times from 5 to 20 min. The optimum volume was chosen at 10 mL for grafting on the cotton fabric due to the highest % of adsorption, fluorescence emission, and gray intensity as shown in As shown in In this work, the extraction solvent was a mixed solvent system comprising acetonitrile (ACN) and ethanol (EtOH). ACN was used as a solvent for preparation of MT solution and EtOH was used to remove unreacted reagents . TherefoThe scheme of polymerization of the MIPs@EDA-CDs/CF by the non-covalent imprinting approach and extraction of template molecules is illustrated in As illustrated in 2O2 to activate the -OH group of the cellulose [SEM analysis of raw cotton fabric revealed fat and spirally twisted ribbon-like fibers with a rough surface from impurities such as pectin and waxes, as shown in ellulose . The SEM−1 due to the -NH2 and -OH functional groups while the treated cotton fabric showed a low intensity at 3333 cm−1 due to only O-H stretching. The absorption bands with a weak peak around 2900 cm−1 indicated the stretching of aliphatic -CH2- and -CH3 groups. Additionally, the absorption peak at 948 cm−1 corresponded to C-H out-of-plane bending at a double bond of MAA. The presence of two significant peaks around 1721 cm−1 and 1149 cm−1 exhibited the existence of EGDMA due to C=O stretching and C-O stretching, respectively. Moreover, the MIPs/MT@EDA-CDs/CF, the NIPs@EDA-CDs/CF, and the MIPs@EDA-CDs/CF revealed a peak at 1635–1653 cm−1 due to the N-H bending of the carbon dots except that the treated cotton fabric did not show any peak around these wavenumbers. These peaks confirmed that the amino group of EDA-CDs was still grafted on the cotton fabric as well as it was washed with desorption solvent. Hence, this proved that the desorption solvent did not interfere with or reduce the fluorescence on the proposed cotton sensor. In addition, the MIPs/MT@EDA-CDs/CF showed a sharp peak at 1636 cm−1 as α, β-unsaturated C=O stretching of the MT.The treated cotton fabric, the MIPs/MT@EDA-CDs/CF, the NIPs@EDA-CDs/CF, and the MIPs@EDA-CDs/CF were characterized using Attenuated Total Reflectance—Fourier Transform Infrared (ATR-FTIR). As shown in The treated cotton fabric, EDA-CDs/CF, MIPs/MT@EDA-CDs/CF, NIPs@EDA-CDs/CF, and MIPs@EDA-CDs/CF were taken under UV radiation and analyzed using RGB fluorescence imaging to confirm the visualized fluorescence emission on the proposed cotton sensor.As shown in The enhancement fluorescence emission of the MIPs@EDA-CDs/CF after imprinting MT in MIP structure could be explained as follows. MT interacted with MAA through formation of hydrogen bonds as MT has H-bond acceptors. Moreover, MT also interacted with amino groups through hydrogen bonding or electrostatics, which can enhance the conjugation degree of amino-passivated EDA-CDs/CF. As a result, the fluorescence emission increased as shown in A smartphone was used to take optical images of the MIPs@EDA-CDs/CF under UVA irradiation inside the detection box at 90° for fluorometric measurement. These fluorescence images were then processed with the ImageJ program coupled with a personal laptop for gray, red, green, and blue measurements. The RGB values from converting the image with the corresponding gray intensity were compared with a blank to obtain ∆ gray intensity. The ∆ gray values were calculated from the blank subtraction using Equation (1). In comparison with the blank, ∆ red, ∆ green, and ∆ blue intensities with the corresponding red, green, and blue values were calculated from Equations (2)–(4).2 = 0.999 because the highest coefficient is equal to 1. The limit of detection (LOD) and the limit of quantification (LOQ) of the ∆ red intensity were 44.4 μg/L and 134 μg/L, respectively. The LOD and LOQ were calculated based on 3.3 × S/b and 10 × S/b, respectively, where b is the slope obtained from the calibration curve, and S is the standard deviation of the response [The color changes on the MIPs@EDA-CDs/CF were not observed by naked-eye detection but could be observed under UVA radiation. A smartphone-based fluorescence ImageJ program was used for quantitative analysis. Under UVA radiation in a detection blacklight box, the MIPs@EDA-CDs/CF exhibited the lowest blue fluorescence emission in the absence of MT c, which response ,59. Cons+, K+, Ca2+, Mg2+, SO42−, Cl−, phenol, nonylphenol, and testosterone were carried out to study the selectivity of the MIPs@EDA-CDs/CF towards MT detection. All interferences caused no significant differences in fluorescence emission of the MIPs@EDA-CDs/CF (0–4.9%) which was less than NIPs@EDA-CDs/CF as the control sensor (10.5–24.6%) for MT detection, as clearly shown in Different ions, organic compounds, and structural analog molecules of the same concentration (500 μg/L) including NaFrom The MIPs@EDA-CDs/CF was tested with 1000 μ/L of MT for reusability and stability. The MIPs/MT@EDA-CDs/CF was then washed with EtOH:ACN (1:4) to remove MT out and tested again with MT to investigate the ability of the cotton sensor for several cycles. As shown in To evaluate the precision of the proposed method, three different concentrations of MT  were analyzed on the same day (intra-day) and three following days (inter-day). The detection was found to be precise at the ppb level, with %RSD of the five replicates ranging from 1.01–5.40% on the intra-day and 1.29–5.87% on the inter-day.The MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF were respectively tested with 100 μg/L, 500 μg/L, and 1000 μg/L. Then, both cotton sensors were taken out and kept air-dried in a dark place. The remaining solutions were then analyzed by a UV–visible spectrophotometer.The amounts of MT binding to the MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF are shown in Furthermore, the imprinting factor measures the imprinted polymer’s strength of interaction with the template molecule. The high imprinting factor value might interact more strongly with the template molecule and have a greater binding site than non-imprinted polymers . It was In this study, the proposed method was applied to determine MT in water and sediment samples and compared with the HPLC analysis. Spiking MT solutions at 200 μg/L, 400 μg/L, and 600 μg/L into both samples was investigated to calculate the percentage of recovery for an accuracy study.As shown in 17 α-methyltestosterone , methacrylic acid , ethylene glycol dimethacrylate , and ethylenediamine  were purchased from Tokyo Chemical Industry Co., Ltd., Japan. Azobisisobutyronitrile (12 wt% in acetone) was purchased from Sigma Aldrich, USA. Citric acid anhydrous  and ethanol  were purchased from Merck, Germany. Acetonitrile  was purchased from Macron and Avantor, USA. Hydrogen peroxide  was purchased from QReC New Zealand, while sodium hydroxide  was purchased from Fisher, India. An SPE cartridge (C18E 500 mg/30 mL) was purchased from Welch, USA. Cotton fabric, 220 threads per inch, was supplied from a local company in Thailand. ex of 353 nm and an emission wavelength λem of 450 nm, whereas a UV–visible spectrophotometer 1800 Double Beam  was used to determine the remaining MT in a solution.The EDA-CDs were synthesized using a domestic microwave . The synthesized EDA-CDs were characterized using the F-2500 fluorescence spectrophotometer  with an excitation wavelength λ−1  for characterization of EDA-CDs, MIPs/MT@EDA-CDs/CF, MIPs@EDA-CDs/CF, and NIPs@EDA-CDs/CF using PIKE instruments and a single refection horizontal ATR accessory (incident angle of 45°).Thermo Fisher Scientific Nicolet 6700, USA, provided ATR-FTIR spectra ranging from 4000–400 cmThe morphological analysis of the cotton fabrics was performed using a scanning electron microscope  obtained from JEOL, Thailand. Furthermore, an energy-dispersive X-ray spectrometer  was used to determine the chemical composition of raw cotton fabric, treated cotton fabric, EDA-CDs/CF, MIPs/MT@EDA-CDs/CF, MIPs@EDA-CDs/CF, and NIPs@EDA-CDs/CF.The confirmation of visualized fluorescence on the cotton sensor, treated cotton fabric, EDA-CDs/CF, MIPs/MT@EDA-CDs/CF, MIPs@EDA-CDs/CF, and NIPs@EDA-CDs/CF was taken under UV radiation using a highly sensitive and robust charge-coupled device imager  for RGB fluorescence imaging. v/v) with a flow rate of 1.0 mL min−1.The comparison of the proposed method was conducted using an HPLC-UV detector  with a RP-C18 column . The temperature of the column was kept constant at 25 °C, and MT was detected at a wavelength of 245 nm. The mobile phase was water–acetonitrile  was used to take an image of the MIPs@EDA-CDs/CF at 90 degrees under UVA radiation. The optical fluorescence images were analyzed using the ImageJ program to measure gray, red, green, and blue intensities. The detection box, smartphone and setup are shown in An amount of 5 g of CA anhydrous was dissolved with 20 mL of deionized water in a 100 mL beaker, and 2.50 mL of EDA was added to the solution. After adding EDA, the reaction produced heat. Then, the mixture solution was cooled by stirring for 10 min before heating in a microwave at 300 w for 8 min (the beaker was covered by a watch glass). After that, the obtained brown EDA-CDs solution was evaporated by a rotary evaporator and completely dried in an oven at 40 °C for 6 h. Finally, the obtained brown EDA-CDs powder was collected and kept in a dark place for further use.w/v NaOH for removing wax and pectin at 95 °C for 2 h. After that, the cotton fabric was washed with deionized water many times to make it neutral. The treated cotton fabric was then soaked in 250 mL of 5% H2O2 for 2 h before being rinsed with 500 mL of deionized water 3 times. Then, the treated cotton fabric was dried in the oven at 105 °C for 5 min and cut into small pieces (2 × 2 cm). After that, a piece of cotton fabric and 10 mL of 1000 mg/L EDA-CDs solution were placed into a 50 mL Erlenmeyer flask and sonicated for 10 min. Finally, the EDA-CDs grafted cotton fabric (EDA-CDs/CF) was taken out and air-dried in a dark place for further use.Cotton fabric (20 × 20 cm) was treated with 600 mL of 5% v/v) by a vortex mixer level 1 for 10 min. The washing procedure was repeated 3 times to completely remove the MT template molecules and unreacted reagents from the MIPs/MT@EDA-CDs/CF. To confirm that no MT was left, the remaining solutions after washing were analyzed with a UV–visible spectrophotometer and the obtained MIPs@EDA-CDs/CF was then subjected to the detection process using the smartphone-based image analysis method. After no MT was detected, the MIPs@EDA-CDs/CF was kept in a brown zip plastic bag in an airtight container before preserving it in a refrigerator for further use. The non-imprinted composite NIPs@EDA-CDs/CF was prepared as a control using an identical procedure without the addition of MT. An amount of 5 mL of 50 mg/L MT and 300 μL of MAA were added to the Erlenmeyer flask and shaken using a shaking incubator for 30 min. Then, 700 μL of EGDMA and 50 μL of AIBN (12% in acetone) were added to the Erlenmeyer flask. The solution was shaken and polymerized using a shaking incubator at 60 °C for 6 h. After that, a piece of EDA-CDs/CF from +, K+, Ca2+, Mg2+, SO42−, Cl−, testosterone, phenol, and nonylphenol), shaken by hand for 5 min and then kept for another 10 min. After that, both MIPs@EDA-CDs/CF and NIPs@EDA-CDs/CF were taken out and kept air-dried in a dark place. The air-dried MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF were then subjected to the detection process the using smartphone-based image analysis method.The MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF were tested with 5 mL of the mixture between 500 μg/L MT and interferences at the concentration of 500 μg/L  to remove the MT and then subjected to the detection and analysis processes. As mentioned above, the experiments were repeated for 12 cycles.The stability of the MIPs@EDA-CDs/CF is also an important indicator concerning whether the material can be used in actual detection. The MIPs@EDA-CDs/CF was kept under normal light, in a dark place, and in a refrigerator (4−8 °C). Then, the materials were subjected to the detection process using the smartphone-based image analysis method every day for a period of use.When evaluating the precision of the proposed method, it was necessary to assess the repeatability. A piece of the MIPs@EDA-CDs/CF was placed into a 30 mL PE bottle containing 5 mL of three concentrations of MT . The PE bottle was shaken by hand for 5 min and kept still for 10 min. Then, the remaining experiments, as previously mentioned, were performed for detection and analysis. Each experiment was repeated with 5 replicates which were analyzed on the same day (intra-day) and three following days (inter-day).0 was the initial concentration, and Ct is the remaining concentration after adsorption (μg/L) of MT. V (L) is the volume of the initial solution and m (g) is the weight of the MIPs@EDA-CDs/CF or the NIPs@EDA-CDs/CF.The adsorption experiment of specificity and selectivity was carried out to evaluate the binding capacity of the MIPs@EDA-CDs/CF while using the NIPs@EDA-CDs/CF as the control. The MIPs@EDA-CDs/CF or the NIPs@EDA-CDs/CF was tested with various MT concentrations of 100 μg/L, 500 μg/L, and 1000 μg/L. After performing the remaining experiments, as previously mentioned in tion (5) ,61,62:(5P (μg/L) is the concentration of MT bound by the MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF and Ct (μg/L) is the concentration of MT remaining in the solution.The specificity of the MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF was calculated using the partition coefficients of selected homologue hormones between polymers and the solution with the same volume and amount of MT as the adsorption experiment. The partition coefficient (K) was determined based on Equation (6) :(6)K=CpCMIP and KNIP were the partition coefficients of MT for the MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF, respectively.The imprinting factor (IF) was used to evaluate the selectivity properties of the MIPs@EDA-CDs/CF and the NIPs@EDA-CDs/CF. In batch rebinding assays, IF is affected by the analyte concentration and the solvent used. IF was carried out using 5 mL of 500 μg/L of MT in acetonitrile. IF was calculated based on Equation (7) ,64:(7)IFWater and sediment samples were collected from Samutprakan province, Thailand, and kept at 4 °C to prevent biodegradability. Therefore, the samples needed to be analyzed for no longer than 48 h, and the sediment sample needed to be dried before analysis. Solid phase extraction (SPE) was usually used to clean up interferences, impurities, and pre-concentration. In this work, SPE (C18E) was used to change the phase from water into acetonitrile. First, the SPE was pre-conditioned with 5 mL of DI water to moisturize the solid phase, and 10 mL of water sample was loaded through the SPE column. After that, 10 mL of acetonitrile was used to elute the MT at a constant flow rate of 1 mL/min. In addition, the sediment sample was dried in an oven at 60 °C for 8 h and then homogenized using a porcelain mortar and sieved using a mesh size of 200 μm. After that, 0.3 g of dried sample was extracted by 10 mL of acetonitrile using a vortex mixer level 1 for 10 min. Finally, the supernatant of the sediment sample was filtered through filter paper (Whatman No. 1). Both water and sediment samples were filtered with a 0.20 μm nylon syringe filter before analyzing with HPLC and compared with the proposed method using the MIPs@EDA-CDs/CF and smartphone-based image analysis process.The original JPG file of the two-dimensional fluorescence images from the smartphone to the personal laptop was transferred using the Telegram application. The intensity function can be used to represent an image in the RGB color model, where color is specified by expressing how much of each of the red, green, and blue components are present in the image; each component ranges from zero to 255. When all components are set to zero, the result is black; otherwise, the result is white if it all sets as 255. This means that the RGB color space may represent 16,777,216 different colors depending on the red, green, and blue values. Nevertheless, the two related realities can bring up concerns: (1) images with the same appearance can have varying pixel values and (2) images with varied appearances can still have the same pixel values . FurtherSimple techniques were used to fabricate the MIPs@EDA-CDs/CF for MT detection. The obtained MIPs@EDA-CDs/CF was examined using FTIR and SEM-EDS to confirm the success of templating MT in the MIPs@EDA-CDs on the cotton fabric’s surface and confirmed the emission of the MIPs@EDA-CDs/CF using RGB fluorescence imaging. The novel imprinting MIPs@EDA-CDs/CF had sufficient capacity and the cavity caused rapid recognition of MT molecules with better selectivity and specificity due to the adsorption capacity and imprinting factor. The fluorescence images obtained from the smartphone were simply analyzed for the gray, red, green, and blue intensity measurement using the ImageJ program for quantitative analysis of MT in water and sediment samples. Furthermore, the proposed cotton sensor exhibited better accuracy and precision and could be used twice. In addition, the ImageJ program provided trustworthy and accurate data compared with the HPLC. The proposed method proved to be convenient and effective for monitoring the MT in water and sediment samples.


P < 0.05). By “ROC curve analysis,” the best critical point was IL-17 in the growth of sarcopenia. The area that comes under ROC (AUROC) value was 0.627 . The ideal threshold value for IL-17 to estimate sarcopenia was 18.5 pg/mL. In the unadjusted model, IL-17 was considerably linked to sarcopenia . After the covariate adjustment observed in the complete adjustment model , this significance still exists. The results of this study suggest a strong relationship between sarcopenia and IL-17. This study will look at IL-17's potential to serve as a key sarcopenia indicator. This trial is registered with ChiCTR2200022590.Aging slows down the mechanisms behind skeletal muscle weakening and mobility. Increases in inflammation brought on by aging may contribute to some characteristics of sarcopenia. As a result of population aging worldwide, sarcopenia, an age-related disease, has become a huge burden on both individuals and society as a whole. The study of the morbidity mechanism and available sarcopenia treatments has received more attention. The inflammatory response may be one of the most important methods behind the pathophysiology of sarcopenia in the aged, according to the background of the study. This anti-inflammatory cytokine inhibits the ability of human monocytes and macrophages to induce inflammation as well as the production of cytokines like IL-6. Here, we investigate the association between sarcopenia and interleukin-17 (IL-17), an inflammatory cytokine in the aged. There were 262 subjects aged 61-90 years who were screened for sarcopenia in Hainan General Hospital. The subjects were divided into 45 males and 60 females aged 65-79 years (average age: 72.00 ± 4.31 years). 105 patients without sarcopenia were randomly selected among 157 participants. It included 50 males and 55 females, aged 61-76 years (mean age: 69.10 ± 4.55 years) as per the standard definition of the Asian Working Group for Sarcopenia (AWGS). The “skeletal muscle index” (SMI), “hand grip strength” (HGS), “gait speed” (GS), “biochemical indexes,” “serum IL-17 level,” nutritional status, and past medical history of the two groups were evaluated and compared. Compared with the participants without sarcopenia, sarcopenia patients had higher average age; less physical exercise; lower total scores of BMI, pre-ALB, IL-17, and SPPB; and a higher proportion of malnutrition risk (all  Population aging has emerged as a top concern of many nations; it is simple and easy to overlook age-related sarcopenia. An indication of aging is chronic inflammation. Sarcopenia is now a frequent condition among the elderly, placing a significant burden on the global healthcare system and society . Age-relThe causes of sarcopenia may be multifaceted including muscle loss , endocriα) was raised in aged patients with sarcopenia [IL-17 and IL-17F form biologically active heterodimers with moderate potency for inducing inflammatory genes when compared to homodimers. Interleukin-17  is one of the cytokines that was cloned in 1993, but it remained unclear throughout the 1990s and early 2000s. Several early investigations suggested that this cytokine was raised in human inflammatory or autoimmune illnesses and was expressed in a noncanonical T helper cell population . The stuAt the Geriatrics Center of Hainan General Hospital in China, between May 2019 and May 2020, a total of 262 elderly people aged between 61 and 90 were assessed as sarcopenia. In line with AWGS [The experimental subjects are those who are under the age of 60; have tumors that are obviously malignant; are bedridden due to severe stroke, heart failure, renal problems, fractures, and other diseases; are subject to severe endocrine diseases and poor management; and have infectious diseases, autoimmune diseases, and a history of mental illness. 2) yields the skeletal muscle index (SMI) [2) and female SMI (5.4 kg/m2) were used to identify low muscle mass [According to the AWGS  definitiex (SMI) . Male SMcle mass .Using a Xiangshan electronic grip tester, grip strength (HGS), the measure of muscle strength was determined . Three times with each hand, the subjects were instructed to use their best effort. Sarcopenia is diagnosed using the maximum HGS. The AWGS consensus standard defines low HGS as males of 28 kg and females of 18 kg .During the 6-meter process, gait speed (m/s) is employed as a gauge of physical performance. Subjects of the experiment were told to walk six meters at their normal pace [Definition criteria for regular exercise: exercised more than thrice a week, more than 30 minutes each time, and during the past 6 months at least. All of these are considered normal as regular exercisesSmoking definition criteria: the World Health Organization defines “continuous or cumulative smoking for 6 months or more in a lifetime” as a smoker. These participants were partitioned into two groups: smokers and nonsmokersDefinition standard of drinking: according to drinking habits, these participants were separated into two categories: drinkers and nondrinkersAfter fasting for at least 6 hours in the morning, 5 mL of cubital venous blood was drawn from each subject. Hemoglobin (Hb) and blood biochemical indicators were calculated soon after blood was drawn. The leftover serum was stored at -80°C for consequent and later testing of IL-17 levels.Using a brief form of the mini nutritional assessment, we assessed the nutritional risk (MNA-SF). Participants were deemed to be at risk of undernourishment if their scores were less than or equal to 11.IL-17 was quantified by flow cytometry  and reagents from Jiangxi Saiji Biotechnology Co., Ltd. according to the requirements of reagent manufacturers with a normal reference interval of 0–20.6 pg/mL.t-test are used, respectively, for categorical and continuous variables. To assess the strength of the connection between IL-17 and sarcopenia, odds ratios (ORs) were created using logistic regression. The strength of the relationship between IL-17 levels and grip strength, SMI, and speed was further examined in the current study using Pearson's correlation coefficients. A multivariate linear regression model with a progressively changed architecture was used to explain the connection between IL-17 and the sarcopenia component. The current study further examined the strength of the relationship between IL-17 levels and grip strength, SMI, and pace using Pearson's correlation coefficients. IL-17 and the sarcopenia components were related, and we used a multivariate linear regression model with a gradually changed design to explain this link. By using the following extended model linear regression, covariate adjustment was examined. The best IL-17 cutoff value for sarcopenia prediction was investigated using receiver operating characteristic (ROC) curve analysis.SPSS is used for the statistical analysis of this study . Indicators for continuous data were the median and interquartile range (IQR). Utilizing frequency counts and percentages, categorical data were recorded. The chi-square test and independent sample P < 0.05).P < 0.001).In comparison to the no sarcopenia group, the grip strength, SMI, and walking speed in the sarcopenia group were lower and the three continuous variables were statistically significant (P = 0.004). After the covariate adjustment observed in model 2  and model 3 , aboriginality persisted.In P = 0.002). The “optimal cutoff value of 18.5 pg/mL” was established by the maximum “Youden's index” with a sensitivity of 0.829 and specificity of 0.381.By “ROC curve analysis,” β = −0.263, 95% CI = −0.101, -0.034, P < 0.001), HGS , and gait speed , showed statistical implication and importance in the unadjusted model. Similar trends were noticed in the fully adjusted model. The β coefficients of SMI, HGS, and gait speed were -0.212 , -0.130 , and -0.169 .This study examines the relationship between IL-17 levels and the type of sarcopenia in representative samples of older sarcopenic patients. Our findings demonstrated a strong correlation between IL-17 levels and sarcopenia. As we know, the role of serum IL-17 in predicting sarcopenia was investigated for the first time in this study. Additionally, the NLRP3 inflammasome plays a role in the reduction in muscle glycolysis that comes with age. Age-related alterations in muscle glycolytic enzyme activity and myofiber size are decreased when NLRP3 is eliminated.α [Studies have stressed the connection between (chronic) systemic inflammation and a decline in skeletal muscle function , as wellα  went up.α . The risα . ER streα .The aged patients will have chronic low-grade inflammation when getting old . Aging iIn older individuals with sarcopenia, the study showed that IL-17 was inversely correlated with grip strength and walking speed. Walking speed and grip strength are indicators of physical function in the elderly. At present, there is no separate research report on IL-17 and walking speed, but Kositsawat et al.  found thThis study was the first to investigate how serum IL-17 affected the likelihood of developing sarcopenia. The results suggested a substantial association between IL-17 and sarcopenia in both men and women. The level of serum IL-17 may be a useful signal to assess the threat of sarcopenia in a clinical setting, even though additional research and investigations are needed to confirm the potential mechanism.Age-related decline in muscle glycolytic capacity is facilitated by the NLRP3 inflammasome. The reduction in the number of glycolytic myofibers and the decreased activity of glycolytic enzymes in muscle with aging are both lessened by the deletion of NLRP3. This study examines the relationship between IL-17 levels and the type of sarcopenia in representative samples of older sarcopenic patients. Our findings demonstrated a strong correlation between IL-17 levels and sarcopenia. The loss in muscle glycolysis ability that comes with aging is facilitated by the NLRP3 inflammasome. Age-related alterations in muscle glycolytic enzyme activity and myofiber size are decreased when NLRP3 is eliminated. The proinflammatory cytokine IL-6, the anti-inflammatory cytokine IL-10, and the IL-6/IL-10 ratios were all found to be increased in senior sarcopenia patients. Sarcopenia was associated with elevated levels of the proinflammatory cytokine IL-6, the anti-inflammatory cytokine IL-10, and IL-6/IL-10 ratios. By increasing autophagy activity to hasten the clearance of immunological mediators, the NOD-like receptor 3 (NLRP3) inflammasome normally fine-tunes the course of the innate immune response it has triggered. The autophagy response is reduced as we age and develop age-related disorders, immunological mediators are still active, and inflammation is prolonged. Additionally, the selection bias in the current study design may result in an incorrect expression of the connection. The patients' cognitive condition, drug use, and dietary health are crucial aspects of sarcopenia, but they are completely ignored. Therefore, the influence of confusion bias cannot be ignored.


The concentration of a single molecule, N-acetylornithine, makes it possible to discriminate between control and diabetes . In addition, receptor operating characteristic curve and principal component analysis for the above-mentioned six molecules yielded significantly (p < 0.001) altered in the aqueous humor of the patient group. In addition, receptor operating characteristic curve and principal component analysis for six compounds yielded cut-off values and remarkable sensitivity, specificity, and segregation ability. The altered level of N-acetylornithine may be due to an increased amount of acetate in diabetes. It is of interest to further investigate whether this alteration is related to the pathogenesis of the disease. The increase in the amino form of pyruvate, alanine, in diabetes is also relevant because it could be a means of reducing the formation of lactate from pyruvate.The composition of the aqueous humor of patients with type 2 diabetes is relevant to understanding the underlying causes of eye-related comorbidities. Information on the composition of aqueous humor in healthy subjects is limited due to the lack of adequate controls. To carry out a metabolomics study, 31 samples of aqueous humor from healthy subjects without ocular pathology, submitted to refractive surgery and seven samples from patients with type 2 diabetes without signs of ocular pathology related to diabetes were used. The level of 25 molecules was significantly ( Diabetes is a disorder of glucose metabolism and handling that can manifest in early childhood type 1 diabetes) or later in life (mainly type 2 diabetes)  diabetes. Type 2 https://www.who.int/news-room/fact-sheets/detail/diabetes, accessed on 9 July 2022). Ocular alterations cause discomfort and significantly reduce the quality of life of diabetic patients; as an example, the prevalence of dry-eye syndrome is >50% in type 2 patients [According to the World Health Organization “Diabetes is a major cause of blindness, kidney failure, heart attacks, stroke and lower limb amputation” .p < 0.001 and log2 fold change (FC) > |1|. Moving thresholds to p < 0.001 and log2 fold change (FC) > |0.4|, five additional molecules were selected: kynurenine, creatinine, dimethylarginines (DMA), hydroxy-butyryl-carnitine , and alanine. Remarkably, the levels of the selected molecules were increased in diabetes.Analysis of data from healthy controls and patients diagnosed with type 2 diabetes led to finding reliable concentrations for 80 molecules  for N-acetylornithine was 1 and the sensitivity and specificity were 1, that is, the measurement of the level of N-acetylornithine in the aqueous humor would allow to distinguish between healthy controls and diabetic patients. The AUC for the other five molecules was high, close to 0.8, with remarkable sensitivity and specificity values ; consequMetabolomics is becoming essential to understanding the composition of the aqueous humor in both health and disease. Alterations in the concentrations of molecular components in the aqueous humor can provide relevant information to understand whether a certain comorbidity  is the consequence of a chronic disease, such as type 2 diabetes.p < 0.001 and log2FC > |1| (FC: fold change), N-acetylornithine is the only molecule that increases more than two-fold (FC = 3.8) in diabetes samples. The median concentration of N-acetylornithine ranges from 0.41 in the control to 1.55 µM in the diabetes samples. When the threshold drops to p < 0.001 and log2FC > |0.4|, there are five other molecules that increase their concentration in diabetes: kynurenine, creatinine, total dimethylamines, hydroxy-butyryl-carnitine, and alanine. In terms of the absolute value of the change, we consider alanine and creatinine to be relevant; their median concentration increased in diabetes to >70 and >15 µM, respectively. Kynurenine increased from 0.45 in the control to 0.65 µM in the diabetes samples.We present here complementary information related to the composition of AH samples from healthy individuals without known eye disease. In a previous study we could analyze only 16 samples from healthy individuals and, in this study, we have incorporated data from 15 more healthy individuals. The median value of all molecules whose concentration can be reliably determined is similar. Adding more controls does not significantly change the findings that were reported in a previous study related to altered AH composition in glaucoma . As in gThrough ROC analysis, the four highest ranked metabolites are BAs . The staIncreased alanine concentration in AH also occurs in glaucoma, but to a lesser extent . The incContrary to what was thought for decades, ocular fluids are not independent of CSF; communication can occur through the optic nerve and the perivascular space of the central retinal artery ,22,23. H1H-NMR-based metabolomic study using AH shows the data consistent with activation of pathways related to energy production and to alanine, glutamate, and aspartate metabolism in patients with diabetes mellitus (and cataracts) [The presence of N-acetylornithine in AH is intriguing because the enzymes that specifically handle this molecule are not expressed in mammals. N-acetylornithine carbamoyltransferase (E.C. 2.1.3.9) uses the molecule to synthesize N-acetyl-L-citrulline, which is then processed to arginine. Additionally, glutamate N-acetyltransferase (E.C. 2.3.1.35) catalyzes the interconversion of glutamate and N-acetyl-L-citrulline to L-ornithine and N-acetyl-glutamate; therefore, N-acetylornithine is relevant in the synthesis of arginine in microorganisms  and ornitaracts) . Additiotaracts)  for reviAlthough a decreased serum creatinine concentration is a risk factor for developing diabetes, once diabetes is established and the kidney has been affected, creatinine levels increase as a consequence of low renal clearance and impaired protein synthesis . IncreasRegarding the over-represented acyl-carnitine in diabetic patients, hydroxy-butyryl-carnitine, it should be that some acyl-carnitines provide benefits in diabetes, from improving insulin sensitivity to controlling oxidative stress ; the committee deemed the study exempt from review. All the patients with diabetes were under good control of glycemia. None of them showed any sign of diabetic retinopathy. The aqueous humor was collected at the beginning of the intervention. In both diabetes and control groups there were individuals with myopia or hyperopia but without cataract or eye pathologies; they underwent surgery for refractive lensectomy or ICL implanting .A total of 38 samples were collected; 31 samples of healthy controls  were collected. None of the patients or healthy controls reported any kidney disease, and creatinine and urea plasma levels were within reference values. Axial length (AXL) was 23.92 mm (21.25–27.29 range) in the control group and 24.37 mm (23.23–25.32 range) in the diabetes group. Glycosylated hemoglobin measured one month before the surgery was within reference values (<40 mmol/mol) in the control group and <45 mmol/mol in the patient group. Three patients had controlled hypertension (43%), one hyperlipidemia (14%), two (29%) a previous episode of myocardial ischemia, and one (14%) did not have any comorbidity. Patient medication for diabetes mellitus was metformin. None of the individuals had previously undergone eye surgery. Participants were informed and, in terms of ethical standards, this study adhered to the tenets of the declaration of Helsinki. The study has been evaluated by the Surgeries were performed in fasting conditions by J.S.N. The pupil was dilated and disinfected with 5% povidone iodine 5. Topical anesthesia was used and the surgery protocol was identical in all cases. The first side port, approximately 1 mm width, was performed using the microscope in the operating room. 100–150 µL AH was aspirated with a 27G needle. Samples were immediately transferred to 0.5 mL Eppendorf tubes and stored at −80 °C until analysis.www.biocrates.com/products/research-products/absoluteidq-p180-kit, (accessed on 16 October 2022). Up to 30 μL of aqueous humor was plated in each well. The initial sample processing was as indicated by the manufacturer. Afterwards, derivatized samples were analyzed in the AB Sciex 6500 QTRAP MS/MS mass spectrometer  coupled to an Agilent 1290 Infinity UHPLC system . Data analysis was performed using Analyst and the MetIDQ™ 5.5 software.The AbsoluteIDQ™ p180 Kit , was used. 188 metabolites from BAs, amino acids, hexoses, phospho- and sphingolipids, and acyl-carnitines can be determined using this specific kit. Individual metabolites may be found in t test, comparing one by one all those molecules whose concentration was reliably determined. Significant differences were considered when p < 0.001. For closer comparisons we selected those metabolites that met both criteria: log2 fold change (FC) > |0.4| and p-value < 0.001. Receiver operating curves (ROC) were made using the IBM SPSS software. Principal component analysis (PCA) was made using the R software.Univariate analysis of data was performed using two-sample


Amphiphilic polysaccharides can be used as wall materials and applied to encapsulate hydrophobic active chemicals; moreover, there is significant demand for novel medical high-molecular-weight materials with various functions. In order to prepare amphiphilic schizophyllan (SPG), octenyl succinic anhydride (OSA) was chosen to synthesize OSA-modified schizophyllan (OSSPG) using an esterified reaction. The modification of OSSPG was demonstrated through FT-IR and thermal analysis. Moreover, it was found that OSSPG has a better capacity for loading curcumin, and the loading amount was 20 μg/mg, which was 2.6 times higher than that of SPG. In addition, a hydrogel made up of PVA, borax, and C-OSSPG (OSSPG loaded with curcumin) was prepared by means of the one-pot method, based on the biological effects of curcumin and the immune-activating properties of SPG. The mechanical properties and biological activity of the hydrogel were investigated. The experimental results show that the dynamic cross-linking of PVA and borax provided the C-OSSPG/BP hydrogel dressing with exceptional self-healing properties, and it was discovered that the C-OSSPG content increased the hydrogel’s swelling and moisturizing properties. In fibroblast cell tests, the cells treated with hydrogel had survival rates of 80% or above. Furthermore, a hydrogel containing C-OSSPG could effectively promote cell migration. Due to the excellent anti-inflammatory properties of curcumin, the hydrogel also significantly reduces the generation of inflammatory factors, such as TNF-α and IL-6, and thus has a potential application as a wound dressing medicinal material. Skin, as the human body’s first line of defense , once daSchizophyllum commune with a 1,3-linked glucan backbone and a 1,6-linked D-glucose side chain at every third residue [Schizophyllan (SPG) is an extracellular, naturally occurring, and high-molecular-weight polysaccharide produced by  residue . SPG has residue , and som residue , migrati residue , and der residue . At the  residue  and mois residue . The pos residue . Thus, t residue . The com residue , and the residue ,24. The  residue , Arabic  residue , and sta residue , which hIn addition, SPG can form a gel in the presence of sorbitol or borax, but the gel is relatively weak and brittle . A combiIn this work, we synthesized an amphiphilic SPG by modifying it with octenyl succinic anhydride. The results of the infrared and thermogravimetric analyses showed that octenyl succinic acid was successfully grafted onto the SPG. The curcumin loading of the OSSPG was 2352 times that of curcumin in an aqueous solution, which provided a new strategy for the solubilization of hydrophobic substances. Meanwhile, a double network hydrogel with self-healing abilities and certain mechanical properties was prepared using the one-pot method. The self-healing, stretching ability, swelling, and water loss of the hydrogel were systematically investigated. In addition, the wound healing of the composite hydrogel dressing was evaluated through in vitro experiments. This work provides new insights into the use of OSSPG as wall materials for encapsulating complex and bioactive components, thus facilitating the development and application of OSSPG as a novel wound-healing material.−1 in the spectrum of SPG originated from the O–H stretching of the hydrogen bonds [−1 and 1640 cm−1 are assigned to the C-H stretching and variable angle vibrations [−1 refers to the C–O linkage. Absorptions at 889 cm−1 are typical for β-D-glucose in a pyranose form [−1 in the spectrum of OSSPG is attributed to the C=O stretching vibration of the carboxylic group of OSA [−1 for OSSPG can be ascribed to the asymmetric stretch vibration of carboxylate (RCOO−) [The FTIR spectra were applied to determine the structural differences between SPG and OSSPG, and the results proved the successful esterification of SPG a. The wien bonds . The peabrations ,34. The ose form . As showp of OSA . The abs (RCOO−) , which p (RCOO−) .−1. The hydroxyl group of curcumin had an absorption at 3510 cm−1 [−1 and the stretching vibrations of the benzene ring showed sharp absorption bands at 1603 cm−1. The characteristic peak at 1283 cm−1 was due to the aromatic C–O stretching vibrations, and the peaks at 856 cm−1 were attributed to the C–O–C stretching vibrations of curcumin [−1 and 1580 cm−1 remained, and the phenolic hydroxyl characteristic peak at 3510 cm−1 and 1630 cm−1 disappeared due to the mask of the signal peak due to the embedding of curcumin into the hydrophobic core formed by the vector. The C-OSSPG did not change significantly compared to the OSSPG profile, indicating that no other chemical bonds were generated, and there were only intermolecular interactions between curcumin and OSSPG.Curcumin is a polyphenolic substance present in an enol structure, with no obvious absorption peak at 1800–1650 cm510 cm−1 . The C=Ocurcumin . The chaAs seen in Curcumin is a water-insoluble polyphenol substance with a solubility of only 17 ng/mL in water. As shown in 0.2/BP before and after hydrogel formation, respectively. 0.2/BP hydrogel exhibited elastic-like behavior [behavior . Additio0.2/BP hydrogel are presented in 0.2/BP hydrogel arising at 832 and 665 cm−1 were assigned to the B-O stretching from the residual B(OH)4− and the bending of the B-O-B linkages within the borate network [−1 after the addition of B molecules within the PVA hydrogels [The FT-IR spectra for PVA, borax, BP, and the C-OSSPG network , respectydrogels . All of 0.2/BP hydrogels. The porous structure of the BP hydrogel is shown in 0.2/BP hydrogel changed. As a result of the intermolecular interaction between OSSPG and PVA, the majority of the pore space in the BP hydrogel was filled with OSSPG, resulting in smaller and denser pores [The study of the surface morphology was performed using SEM to detect the cross-linking pattern of the BP and C-OSSPGer pores .0.2/BP hydrogel. Two colored 6 cm columnar-shaped hydrogels were cut into two pieces and then recombined at the fracture surfaces. Two pieces of fractured hydrogel healed autonomously and rapidly after 1 min of self-healing, without any external intervention, at room temperature. This is due to the presence of dynamic boronic ester linkages in the hydrogel that can satisfy the rapid healing of the hydrogel [hydrogel . Moreove0.2/BP hydrogel can reach six times its weight, more than 1.3 times that of the BP hydrogel. The reason for this might be that OSSPG has abundant hydrophilic groups [0.2/BP hydrogel ranks this material as a promising wound dressing.The influence of different hydrogels on the swelling performance was analyzed. c groups , such as0.2/BP hydrogel has good water retention. It is ascertained that PVA can retain a certain amount of water, and the addition of C-OSSPG increased the number of hydrophilic hydroxyl groups, which can be more closely combined with water for hydrogels.Maintaining a moist environment is also critical for wound healing. Lightly moist environments can promote wound healing , and hydCytotoxic studies are needed to avoid toxicity in normal cells. Wound healing is a complex physiological process that requires the synergy of multiple cells during healing . Keratin0.2/BP, but promoted the healing of the wound without any significant differences compared to the control group. After 48 h, the C-OSSPG0.2/BP and C-OSSPG0.15/BP hydrogels promoted the in vitro wound closure rate significantly (p < 0.01). This shows that the hydrogels containing C-OSSPG at concentrations of 0.2 wt% and 0.15 wt% had excellent cell-healing-promoting effects on the BJ cells, thus accelerating wound healing. However, wound healing is a complex process, so further investigation may be required before its application to clinical use. Considering the appropriate water storage capacity, excellent water absorption, and cell migration ability, the C-OSSPG0.2/BP hydrogel was chosen to further study their anti-inflammatory properties.Fibroblasts not only play a pulling role in the wound healing process, but part of them can also differentiate into myofibroblasts, which can secrete ECM and better promote wound healing . Figure 0.2/BP hydrogel, although they increased by 16.5% after being treated with the BP hydrogel. This suggests that the use of the hydrogel without C-OSSPG alone causes further inflammation. Similar to TNF-α, the C-OSSPG0.2/BP hydrogel can effectively inhibit the production of IL-6 (p < 0.01); the IL-6 contents released from the RAW 264.7 cells reduced by 81.7% after being treated with the C-OSSPG0.2/BP hydrogel  was supplied by Aladdin Biochemical Technology Co., Ltd. . High-glucose DMEM medium and trypsin were provided by U.S. Gibico Corporation. Fetal bovine serum was provided by Hangzhou Sijiqing Bioengineering Materials Co., Ltd. . All the other chemical materials were of minimum analytical grade. The enzyme-linked immunosorbent assay (ELISA) kit for tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) was supplied by Neobioscience Technology Company .Curcumin (≥95.0% purity) was purchased from Sigma-Aldrich Corp.  and 2-Octen-1-ylsuccinic anhydride (99.0% purity) was purchased from Shanghai yuanye Bio-Technology Co., Ltd. . Schizophyllan was obtained from the fermentation products of w/v) was gradually added to the obtained liquid supernatant for the precipitation of crude polysaccharides. After standing for 12 h at 4 °C, the precipitated SPG was collected through centrifugation and finally freeze-dried.The SPG was prepared by the method described by Deng et al. . The finThe OSSPG was synthesized according to a previously reported method for starch octenyl succinate, with slight modifications . Precise2 = 0.9991, where y is the OSA content (mg/mL), and x is the peak area of trans-2-OSA in the liquid chromatogram. The substitution of the final modified polysaccharide was 0.023.The substitution degree (DS) of OSSPG was measured using the HPLC method, as described by Qiu et al. . The staThen, 10 mg of curcumin was added to a 50 mL centrifuge tube containing 10 mL of OSSPG at a concentration of 0.2%. Then, the solution was homogenized at 10,000 rpm for 5 min using a homogenizer  to make full contact between the curcumin and the modified polysaccharide. Finally, the solution was centrifuged at 10,000 rpm for 10 min to remove the free curcumin, and the supernatant was collected for further analysis. The filtrate containing the polymeric solution was freeze-dried and stored in the dark at 4 °C for characterization. The final sample was named C-OSPPG, where C signifies the presence of curcumin.2 = 0.9990).The loading capacity (LC) of the curcumin was detected based on a previously published method . A doubl0.05/PB, C-OSSPG0.1/PB, C-OSSPG0.15/PB, and C-OSSPG0.2/PB, respectively.The hydrogels were synthesized using a one-step method, described by Siqi Huang . The hyd−1 with a resolution of 4.0 cm−1. The samples were dried at 105 °C for 12 h before analysis to avoid the interference of moisture.FT-IR analysis: The chemical structures of the SPG, OSSPG, and C-OSSPG were investigated using an FT-IR spectrometer . The spectra were recorded in the wavelength range of 500–4000 cmTG analysis: The thermal stability of the SPG, OSSPG, C-OSSPG, and curcumin were analyzed using a TGA-851e . The samples with a mass between 3 and 5 mg were placed in an alumina crucible at a detection temperature of 50–600 °C, a nitrogen flow rate of 20 mL/min, and a heating rate of 10 °C/min, using nitrogen as a protective gas. An empty pan, sealed in the same manner, was used as a reference.0.2/BP hydrogels were performed using a scanning electron microscope after vacuum-coating the samples with plat. A small piece of dried hydrogel was fixed onto the SEM stubs and sputter-coated with a thin layer of Au prior to examination.SEM analysis: The morphological investigations of BP and the C-OSSPGRheological measurements: The rheological behaviors of the C-OSSPG/BP hydrogels were analyzed with a modular rheometer. The sample was made into a cylindrical shape, with a diameter of 15 mm and a thickness of 6 mm. The temperature (t) was increased at T = 25–65 °C, and the scanning strain (γ) = 0.5%, 0.5 rad/s, to determine the change in the storage modulus and the loss modulus of the sample due to the temperature.0.2/BP hydrogel with a diameter of 10 mm and a length of 60 mm was prepared in advance. To intuitively show the self-healing effect, two C-OSSPG0.2/BP hydrogels were stained with methyl blue and Congo red, respectively. The hydrogel was cut into two pieces, and then the different colors of the two pieces were re-spliced together and stored at 25 °C for 1 min to allow them to self-heal into a complete hydrogel. The tensile capacity of the hydrogel was also tested by a simple stretching of the re-healed hydrogel. Photographs of the hydrogels under different situations were taken.The C-OSSPGd is the weight of the dried hydrogel and Ws is the weight of the swollen hydrogel.The swelling properties of the freeze-dried hydrogel samples were evaluated. Then, all of the samples were immersed in PBS buffer (pH~7) at 37 °C until the gel reached the equilibrium state of swelling. The hydrogel was removed, and the excess solvent on the surface was removed by blotting quickly with absorbent paper, and then weighed. The swelling ability of the hydrogel can be formulated using a swelling ratio; the swelling ratio was calculated as follows:s, Wm, and Wf are the starting weight, measured weight, and last weight of the hydrogel, respectively.The prepared hydrogel was immersed in distilled water. After it reached swelling equilibrium, the hydrogel was placed in an incubator at 50 °C for 24 h under a humid atmosphere of 50%. The weight of the hydrogel was measured at regular intervals until completely dry. The water evaporation rate of the hydrogel can be formulated using the water retention equation as follows:5 cells/mL in 96-well plates, at 100 μL per well. Before the experiments, various synthesized hydrogels were sterilized using UV radiation for 120 min. Subsequently, hydrogel-extracted liquid solutions were prepared by soaking the hydrogels (1 g) in DMEM (4 mL) containing serum at 37 °C for 24 h. The culture media were aspirated, then all of the cells were incubated with extracting liquid solutions for the different hydrogels for 24 h. Finally, the treatment solution was aspirated, and 100 μL of 10% CCK8 solution was added to every well and then incubated in the cell incubator for half an hour before measuring the absorbance value at 450 nm. Cytotoxicity was calculated as the percentage of absorbance in wells with solution-treated cells to that of the control cells (100%), as follows:The cytotoxicity of the hydrogels was evaluated using the ISO 10993-5 method . The in 5/well onto a coated 24-well plate in 10% FBS-DMEM at 37 °C and 5% CO2 to obtain a monolayer of cells. Then, a sterile gun tip was used to draw a line through the monolayer of cells, and then they were washed with PBS. The cells were incubated with the treatment solutions for the different hydrogels or 2% FBS-DMEM (untreated control) for 48 h. The migration rates were observed and photographed under microscopy. The percentage of the closed area was analyzed by using ImageJ, and then compared with the control.The ability of BJ to migrate to the wound environment from adjacent areas determines the speed of wound healing. The wound-healing potential of the hydrogels was assessed using an in vitro wound-healing assay . For thi5 cells/well for 24 h. After removing the cell culture medium, the cells were then stimulated with LPS at 1 µg/mL, followed by treatment with the treatment solutions for the different hydrogels. After incubation for 24 h, the culture medium was collected for the determination of TNF-α and IL-6 levels. The levels of TNF-α and IL-6 were determined using a mouse enzyme-linked immunosorbent assay (ELISA) kit according to the manufacturer’s instructions. The TNF-α and IL-6 results for each sample were calculated from their respective standard curves.RAW 264.7 cells were seeded in 24-well plates at a density of 4 × 10p < 0.05 by Duncan’s multiple range tests. In the chart, * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001.All of the tests were performed in triplicate, unless otherwise indicated, and the values are expressed as the mean ± standard deviation. One-way analysis of variance was performed to evaluate the significance at